Recently, changes in optical microscopy have taken place, providing a possibility of sub-diffraction limited fluorescence imaging, also termed “nanoscopy.” (See Hell S. W., “Toward fluorescence nanoscopy”, Nat. Biotechnol. 2003, 21:1347-55). These advances can be referred to PALM/STORM (see Betzig E. et al., “Imaging intracellular fluorescent proteins at nanometer resolution”, Science 2006, 313:1642-5; and Rust M. J. et al., “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)”, Nat Methods, 2006, 3:793-5) and STED (see Westphal V., “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement”, Science 2008, 320:246-9; and Willig K. I. et al., “STED microscopy reveals that synaptotagmin remains clustered after synaptic vesicle exocytosis”, Nature 2006, 440:935-9).
The concept of PALM/STORM is to repeatedly photoactivate sparse fluorophores with a separation that is greater than the diffraction limit and precisely resolve their locations using a Gaussian fitting procedure. The STED concept operates by depleting the excitable fluorophores surrounding the center of the imaging spot using a donut-shaped beam. Both techniques have provided images of sub-cellular detail with resolutions approaching 30 nm, heretofore only observable by electron microscopy. (See Westphal V. et al., “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement”, Science 2008, 320:246-9; and Huang B. et al., “Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy”. Science 2008, 319:810-3).
The above-described technologies have limitations. For example, the PALM/STORM procedure likely requires the excitation of rare events and currently takes many hours to achieve adequate signal to noise, prohibiting the imaging of living organisms. The STED procedures can work faster (see Westphal V. et al, “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement”, Science 2008, 320:246-9), but may rely on the integrity of a donut beam to populate excited states. The lack of such integrity can limit or prevent sub-diffraction limited imaging deep into tissues, as aberrations in tissue likely destroy the shape of the donut beam. A technique capable of providing sub-diffraction limited imaging of intact or living tissues would likely provide a significant number of possibilities for nanoscopy in the biological sciences. When applied to problems in human medicine, for example, deep tissue nanoscopy in animal and human studies can provide an improved understanding of the molecular mechanisms of tissue issues.
Accordingly, exemplary systems, methods and computer-accessible medium providing sub-diffraction limited imaging of intact or living tissues may be beneficial to overcome at least some of the above-described issues and/or deficiencies.